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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20226653 - 07/05/14 12:28 AM (10 years, 6 months ago) |
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Well....as kizzle asked, did you look beneath the agar?
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: azur]
#20226727 - 07/05/14 12:45 AM (10 years, 6 months ago) |
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A few jars, my wedges ended up against glass and the non colonized side looked fine but at that point it had been shaken so there could have been bacteria that got rubbed off.
But I always look at my plates from the underside as well with a light shining through kinda like candling an egg, just to make sure there's not something I don't see on top.
Also if there was bacteria, it wouldn't account for the issues I'm having.
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20226757 - 07/05/14 12:53 AM (10 years, 6 months ago) |
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How long are you soaking your grain? What kind of grain are you using?
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: azur]
#20226796 - 07/05/14 01:01 AM (10 years, 6 months ago) |
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Wbs, and up to 24 hours depending on hydration.
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20226812 - 07/05/14 01:05 AM (10 years, 6 months ago) |
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I've only used wbs once and didn't like it. Too clumpy. I'm stumped now. Sounds like it's gotta be in your agar or in the air when transferring your agar to grain. Someone with wbs experience chime in please.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: azur]
#20226832 - 07/05/14 01:13 AM (10 years, 6 months ago) |
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I doubt it has anything to do with the grain prep unless somehow my pc is defunct.
As far as being in the air or my agar, its possible but I'm doubting it because I never see it on any plates when I'm doing transfers and isolations.
I've got several pc runs to do tomm so I'll flip my lids and have a go at that. Maybe it is just as simple as my jars not sealing well enough. Especially since I "punch" holes in my lids instead of drilling.
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blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Where are my contams coming from? [Re: Ghatti]
#20226842 - 07/05/14 01:16 AM (10 years, 6 months ago) |
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insert twilight zone emoticon
try taping the area btw lid and jar with masking tape. i do that with LC's/plasictic lids, and it helps a lot. never had to do it with grain tho edit: i do it with parafilm, but i'd imagine masking tape, electric tape, etc would do the same?
-------------------- A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!
The whole is greater than the sum of its parts
Edited by blindingleaf (07/05/14 01:17 AM)
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StygianKnight
A Mushroom

Registered: 03/12/12
Posts: 2,717
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Re: Where are my contams coming from? [Re: blindingleaf]
#20226877 - 07/05/14 01:34 AM (10 years, 6 months ago) |
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Again, do control jars, it's the only way to not just stumble around in the dark guessing.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: StygianKnight]
#20229423 - 07/05/14 07:32 PM (10 years, 6 months ago) |
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Did a pc run earlier and left 2 control jars laying around. Also pc'd my sleeves and righted my lids (gasket down). If I still get a contam I'm likely to jump straight to my agar but I fail to reason why I can't see any contams on it.
Again I don't believe it to be my aseptic technique as none of my agar work gets visibly contamed save for a plate here and there that winds up with a spot or two of bacteria.
But.....here's a question. If you have a jar that had a contam in it and you empty said jar and use it for agar, counting on a pc run to sterilize your agar/jar, how long would you go for?
I've been pc'ing my agar 45mins.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: Ghatti]
#20229459 - 07/05/14 07:45 PM (10 years, 6 months ago) |
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Just wash the jar with dish soap and use it like normal. I PC my agar for 35 mins @ 15psi.
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20229469 - 07/05/14 07:48 PM (10 years, 6 months ago) |
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I always wash containers even before pcing. I guess the time depends the contamination type
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: azur]
#20229515 - 07/05/14 07:58 PM (10 years, 6 months ago) |
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Well we will see what happens. Like I said if it happens again I'll blame dirty specimen on agar but I don't see how.
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20229665 - 07/05/14 08:39 PM (10 years, 6 months ago) |
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What is the said contam 4 posts up?
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: azur]
#20229803 - 07/05/14 09:10 PM (10 years, 6 months ago) |
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Ah, cobweb.
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: Where are my contams coming from? [Re: Ghatti]
#20229881 - 07/05/14 09:26 PM (10 years, 6 months ago) |
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TMC says spores of dactylium dendroides are killed when exposed to 115-122°f for only 30 min. I think it's your air quality. Time to do a serious cleaning and build a flow hood.
How are you sterilizing any tools you are using to transfer? And how pure is your strain?
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blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Where are my contams coming from? [Re: azur]
#20229913 - 07/05/14 09:33 PM (10 years, 6 months ago) |
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OP i really wish u luck with this whole thing, but i just realized, there are no pics of anything from you, agar, grains, bulk, casings, nada. if i were u, i'd take as many pics as possible to try and figure this out!! maybe one of the thousands of posters on this board will recognize something you/we don't by ur text descriptions. sometimes con tams are tricky man, a text description can only go so far.
-------------------- A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!
The whole is greater than the sum of its parts
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i GrOw StUFF
Stranger

Registered: 01/14/11
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Last seen: 10 years, 4 months
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Re: Where are my contams coming from? [Re: blindingleaf]
#20230108 - 07/05/14 10:13 PM (10 years, 6 months ago) |
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If it was bacteria I may question the PC, but it's mold. Even if you botched up the PC process, odds are the mold is killed since it is destroyed at very low temps. Rule out the PC. First thing I would question/change is polyfil for jars GE. Polyfil for a jars GE is outdated IMO. Spend the small amount of cash on mycology grade synthetic filter discs, they can be used tons of times and you won't ever have to question the filter again. How are you making your transfers? In a glove box or in front of a flowhood? 2nd thing I'd question in the transfer/sterile technique.
-------------------- The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Where are my contams coming from? [Re: i GrOw StUFF]
#20230134 - 07/05/14 10:18 PM (10 years, 6 months ago) |
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I use poly and I never question my filters. I even reuse the same poly for a few runs.
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i GrOw StUFF
Stranger

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Re: Where are my contams coming from? [Re: MudaFuka]
#20230169 - 07/05/14 10:24 PM (10 years, 6 months ago) |
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Sweet, all preference. Synthetic discs take the guessing game out though.
-------------------- The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: Where are my contams coming from? [Re: i GrOw StUFF]
#20230216 - 07/05/14 10:31 PM (10 years, 6 months ago) |
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I flame sterilize my scalpel red hot, move inside my sab, quench on a plate, then transfer. BL I am aware about the pics or lack there of, but the only pic I could post that would even be remotely helpful would be of my agar plates. My jars, lids, pc, etc look just like every other one on the forums.
I know sfd's are ultimately superior but using poly should not cause any issues, too many of us here use them without incident.
I work in a still room, in a wet sab, use tyvek sleeves and gloves and usually throw on a mask when I'm going to be in the box for more than just a minute. I'm quick but deliberate and try to limit movements and their impact. I really don't see how I could improve my technique without owning a hood and even then its not my technique that would change but my environment.
I'm really leaning towards my lids somehow but time will tell I suppose. If the batch I did today go bad, then I only have my agar or myself to blame next.
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